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Infection and Immunity, February 1999, p. 653-658, Vol. 67, No. 2
0019-9567/99/$00.00+0

An In Vitro Tissue Culture Bilayer Model To Examine Early Events in Mycobacterium tuberculosis Infection

K. A. Birkness,1 M. Deslauriers,1 J. H. Bartlett,2 E. H. White,2 C. H. King,3 and F. D. Quinn1,*

Division of AIDS, STD and TB Laboratory Research1 and Division of Viral and Rickettsial Diseases,2 National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333, and Division of Infectious Diseases, Emory University School of Medicine, Atlanta, Georgia 303223

Received 26 August 1998/Returned for modification 17 September 1998/Accepted 30 November 1998

A tissue culture bilayer system that mimics some aspects of early alveolar infection by Mycobacterium tuberculosis was developed. This model incorporates human lung epithelial type II pneumocyte (A549) (upper chamber) and endothelial cell (lower chamber) layers separated by a microporous membrane. This construction makes it possible to observe and quantify the passage of bacteria through the two layers, to observe the interaction of the bacteria with the various cell types, and to examine the basic mechanisms of immune cell recruitment to the site of infection. After 107 organisms were added to the upper chamber we microscopically observed large numbers of bacteria attached to and within the pneumocytes and we determined by viable-cell counting that a small percentage of the inoculum (0.02 to 0.43%) passed through the bilayer into the lower chamber. When peripheral blood mononuclear cells were added to the lower chamber, microscopic examination indicated a migration of the mononuclear cells through the bilayer to the apical surface, where they were seen associated with the mycobacteria on the pneumocytes. The added complexity of the bilayer system offers an opportunity to define more precisely the roles of the various lung cell types in the pathogenesis of early tuberculosis.

* Corresponding author. Mailing address: Bldg. 5, Rm. B38, M/S G11, Centers for Disease Control and Prevention, Atlanta, GA 30333. Phone: (404) 639-3205. Fax: (404) 639-4192. E-mail: fdq1{at}cdc.gov.

Infection and Immunity, February 1999, p. 653-658, Vol. 67, No. 2
0019-9567/99/$00.00+0


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